Figure 3. DNAPK regulates Wnt signaling pathway.

A, Scatter plot represents the GSEA normalized enrichment scores (NES) for all pathways associated with DNAPK in both the in vitro knockdown experiment (using VCaP, C4-2B, PC-3, DU145 cells) as well as in the clinical samples from the Mayo Clinic. The Wnt pathway had the top average NES, with high scores in both the clinical and in vitro knockdown results, and is shown in red. Dots in black denote other pathways. B, the bar graph shows the expression of select Wnt pathway genes in LNCaP-AR and C4-2B (CRPC) cells compared to LNCaP (hormone-naive) cells. GAPDH was used as an internal reference (N=3 in duplicates). C, the growth curves show the effect of androgen depletion on proliferation of LNCaP cells, compared to LNCaP cells grown under normal serum (N.S.) conditions (N=3 in duplicates). D Bar graphs represent the expression of Wnt genes in LNCaP cells grown under androgen-depleted (charcoal-stripped serum) conditions, LNCaP cells grown under normal serum conditions, and after DNAPK knockdown in LNCaP cells grown under charcoal-stripped serum conditions, compared to LNCaP cells grown under charcoal-stripped conditions. (N=3, in duplicates). E, Cell growth curves demonstrate the effect of DNAPK knockdown on proliferation of LNCaP cells that developed ADT resistance (from C) (N=2 in duplicates). A subset of these cells was switched to normal serum conditions as a reference. *p < 0.05, **p < 0.001. P-values were calculated using students t-test (Figure 3B, D) or two-way ANOVA (Figure 3C, 3E). All data are represented as mean ± S.D. RQ = relative quantity.